For better understanding the chemical or biological information of ZNF191 （243-368）, we expressed the fusion protein of GST and ZNF191 （243-368）, and used it to obtain the binding DNA sequence of this zinc finger ...For better understanding the chemical or biological information of ZNF191 （243-368）, we expressed the fusion protein of GST and ZNF191 （243-368）, and used it to obtain the binding DNA sequence of this zinc finger protein. But in the process of expression and purification, we found this fusion protein slowly degradated. For resolving this problem, we simultaneously added charged amino acids L-Arg and L-Glu to the solution of fusion protein, and demonstrated that this method can dramatically increase the stability of this fusion protein. This method can make the fusion protein suitable for the continuous works, especially for situations where high protein concentration and long-term stability without precipitate and degradation of protein are required.展开更多
Two mutants of the zinc finger protein, ZNF191(243—368) I323W and R327W, are suc-cessfully obtained by site-directed mutagenesis. The fluorescence spectrum is used to study the interaction between these two mutants a...Two mutants of the zinc finger protein, ZNF191(243—368) I323W and R327W, are suc-cessfully obtained by site-directed mutagenesis. The fluorescence spectrum is used to study the interaction between these two mutants and the oligonucleotides. The influence of the mutation on the interaction has been studied using ethidium bromide (EB) as the fluorescence probe. The binding constants of the I323W-DNA and R327W-DNA have been calculated and the possible binding models have been discussed.展开更多
文摘For better understanding the chemical or biological information of ZNF191 （243-368）, we expressed the fusion protein of GST and ZNF191 （243-368）, and used it to obtain the binding DNA sequence of this zinc finger protein. But in the process of expression and purification, we found this fusion protein slowly degradated. For resolving this problem, we simultaneously added charged amino acids L-Arg and L-Glu to the solution of fusion protein, and demonstrated that this method can dramatically increase the stability of this fusion protein. This method can make the fusion protein suitable for the continuous works, especially for situations where high protein concentration and long-term stability without precipitate and degradation of protein are required.
基金This work was supported by the National Natural Science Foundation of China (Grant Nos. 30170228 and 39990600).
文摘Two mutants of the zinc finger protein, ZNF191(243—368) I323W and R327W, are suc-cessfully obtained by site-directed mutagenesis. The fluorescence spectrum is used to study the interaction between these two mutants and the oligonucleotides. The influence of the mutation on the interaction has been studied using ethidium bromide (EB) as the fluorescence probe. The binding constants of the I323W-DNA and R327W-DNA have been calculated and the possible binding models have been discussed.
